钙荧光白 (Calcofluor white; CFW) 又称荧光增白剂，是一种非特异性荧光增亮染料。真菌 (如藻类等) 细胞壁上含有较多的几丁质及纤维素等β-多糖类物质，可与荧光增白剂中的荧光素结合形成复合物，在荧光显微镜下可看到特定激发光下产生的荧光。在植物生物学中，钙荧光白常用于显示藻类和高等植物中细胞壁生物合成过程中细胞壁的生成变化，以及原生质体细胞的鉴定。
真菌荧光染色液 (一步法) 主要由荧光增白剂、伊文思蓝、氢氧化钾、二甲基亚砜、丙三醇及去离子水组成，其中荧光增白剂可与多糖物质结合产生荧光，伊文思蓝可抑制背景荧光着色，氢氧化钾可溶解角质，有助于更清晰的观察真菌结构，丙三醇有一定的抗荧光淬灭功能。作为一种快速检测真菌的新型染色液，本品可用于各种疑似真菌的荧光染色检查，如手足藓、花斑藓和肺部真菌感染等，在荧光显微镜下快速定性检测真菌。此外，本产品还能检测出卡氏肺孢子虫囊肿，寄生物(例如疟原虫)，以及各类地域性变异真菌菌丝角蛋白、胶原蛋白、弹性纤维也能使用本品进行检测，可将这些物质的结构边界进行荧光标记从而能够明显区分并检出。该试剂仅用于科研领域，不适用于临床诊断或其他用途。
自备材料：
1、新鲜或冰冻临床样本，石蜡及乙二醇甲基丙烯酸酯包埋的组织样本。
2、载玻片、盖玻片、荧光显微镜
操作步骤 (仅供参考)：
1、直接向样本中滴加1滴染色液，以覆盖或淹没整个标本为准，保持染色2分钟。
2、盖上盖玻片，使用棉签或纸巾压片，使标本尽量压成薄片，吸去多余染色液。
3、置于荧光显微镜 (激发波长为340nm~380nm，发射波长为440nm) 下直接观察结果。
染色结果：
样本在荧光显微镜下呈现强或弱蓝绿色荧光，且形态学可辨认，则判断样本为阳性；
若形态学不可辨认、极弱荧光或无荧光，则判断样本为阴性。
注意事项：
1、实验前应做好个人防护，检测样本存在潜在感染性。
2、试剂每次使用时，应及时盖好保存，防止污染，以免影响染色效果。
3、污染废弃物按规定处理，必须符合国家环境和健康安全规定。
4、使用前请详细阅读使用说明书，在有效期内使用。
5、试剂贮存时要尽量避免高温及低温环境，切勿冷冻，同时不能阳光直射。
6、为了您的安全和健康，请穿实验服并戴一次性手套操作。
7、试剂开封后请尽快使用，以防影响后续实验效果。
Calcofluor White (CFW), also known as fluorescent brightener, is a non-specific fluorescent brightening dye. The cell walls of fungi (such as algae) are rich in β-polysaccharides including chitin and cellulose, which can bind to the fluorochrome in the fluorescent brightener to form complexes. Under a fluorescence microscope, specific fluorescence can be observed when excited by a corresponding excitation light source. In plant biology, Calcofluor White is often used to visualize the changes in cell wall formation during the cell wall biosynthesis of algae and higher plants, as well as the identification of protoplast cells.
Fungal Fluorescence Staining Solution (One Step) is mainly composed of fluorescent brightener, Evans blue, potassium hydroxide, dimethyl sulfoxide (DMSO), glycerol, and deionized water. Among these components, the fluorescent brightener can bind to polysaccharides to produce fluorescence; Evans blue can suppress the background fluorescence staining; potassium hydroxide can dissolve cutin, facilitating clearer observation of fungal structures; glycerol has a certain anti-fluorescence quenching effect. As a novel rapid fungal detection stain, this product can be used for fluorescent staining examination of various suspected fungal infections, such as tinea manuum, tinea pedis, tinea versicolor, and pulmonary fungal infections, enabling rapid qualitative detection of fungi under a fluorescence microscope. In addition, this product can also detect Pneumocystis carinii cysts, parasites (e.g., Plasmodium), as well as various bacteria. It can fluorescently label the structural boundaries of these substances, allowing for clear differentiation and detection. This reagent is intended for research use only and is not suitable for clinical diagnosis or any other purposes.
Materials to Be Prepared by User:
1. Fresh or frozen clinical samples; tissue samples embedded in paraffin or glycol methacrylate (GMA)
2. Glass slides, cover slips, fluorescence microscope
Operating Procedures (For Reference Only):
1. Add 1 drop of the staining solution directly onto the sample, ensuring the entire specimen is covered or submerged. Incubate for 2 minutes.
2. Place a cover slip over the sample. Press gently with a cotton swab or absorbent paper to flatten the specimen into a thin layer, then absorb the excess staining solution.
3. Observe the results directly under a fluorescence microscope with an excitation wavelength of 340 nm–380 nm and an emission wavelength of 440 nm.
Staining Results:
Positive: The sample exhibits strong or weak blue-green fluorescence under the fluorescence microscope, and the morphological features are identifiable.
Negative: The morphological features are unidentifiable, or the sample shows extremely weak fluorescence or no fluorescence at all.
Precautions:
1. Personal protective measures should be taken prior to the experiment, as the test samples may carry potential infectivity.
2. Always recap the reagent container immediately after each use to prevent contamination, which could otherwise compromise the staining results.
3. Contaminated waste materials must be disposed of in accordance with national environmental and health safety regulations.
4. Please read the instruction manual thoroughly before use, and make sure to use the reagent within its expiration date.
5. Store the reagent away from extreme high or low temperatures; do not freeze it, and avoid direct sunlight exposure.
6. For your safety and health, wear a lab coat and disposable gloves during the operation.
7. Use the reagent as soon as possible after opening to avoid affecting the outcomes of subsequent experiments.
ZJ26F0838319
危险品化学品经营许可证（带存储）